Bio-prospective potential of Pleurotus djamor and Pleurotus florida mycelial extracts towards Gram positive and Gram negative microbial pathogens causing infectious disease.

BACKGROUND: The emergence of resistance to commonly used antibiotics by human infections occurred mostly due to their overuse, that prompted individuals to pursue novel and innovative treatments. The phytochemical characteristics, antibacterial activity, and cytotoxicity of MCF7 cells were evaluated in two Pleurotus spp. mycelial extracts in this work. METHODS: Pleurotus djamor and Pleurotus florida mycelial extracts from pure cultures were tested for antibacterial activity by a well-diffusion assay and antimicrobial activity against mold fungi was evaluated for biomass inhibition. Mycelial extracts were obtained from dichloromethane extracts and their biophysical characteristics are analyzed by UV-vis spectrum and FTIR analysis. By spraying detection reagents onto TLC plates, the chemicals in dichloromethane extraction of chosen mushroom fungus mycelia were identified. Using the MTT test, the cytotoxic effect of dichloromethane extracts of selected mushroom fungi was evaluated on MCF7 Cell lines. RESULTS: Mycelial extracts of P. djamor and P. florida exhibited significant antimicrobial effect on the bacterial and fungal pathogens tested. Dichloromethane mycelial extracts were obtained using soxhlet extraction which response positive for various phytochemical analysis. Detection of metabolites in thin layer chromatography using spray reagents documented one of few first accounts on flavonoids, anthroquinone and terpenoid compounds in P. djamor and P. florida. P. djamor and P. florida had dose-dependent antiproliferative activity against MCF7 cells, with an inhibitory impact of 55.72% and 64.47% percent at 125 µg/mL, respectively. CONCLUSION: The study has reported the identification with the potent biological activity of some of the key bioactive components present in DCM extracts from the mycelia of P. djamor and P. florida.

Production, partial purification and characterization of ligninolytic enzymes from selected basidiomycetes mushroom fungi.

In recent years, many research on the quantity of lignocellulosic waste have been developed. The production, partial purification, and characterisation of ligninolytic enzymes from various fungi are described in this work. On the 21st day of incubation in Potato Dextrose (PD) broth, Hypsizygus ulmarius developed the most laccase (14.83 × 10-6 IU/ml) and manganese peroxidase (24.11 × 10-6 IU/ml), while Pleurotus florida produced the most lignin peroxidase (19.56 × -6 IU/ml). Laccase (Lac), lignin peroxidase (LiP), and manganese peroxidase (MnP), all generated by selected basidiomycetes mushroom fungi, were largely isolated using ammonium sulphate precipitation followed by dialysis. Laccase, lignin peroxidase, and manganese peroxidase purification findings indicated 1.83, 2.13, and 1.77 fold purity enhancements, respectively. Specific activity of purified laccase enzyme preparations ranged from 305.80 to 376.85 IU/mg, purified lignin peroxidase from 258.51 to 336.95 IU/mg, and purified manganese peroxidase from 253.45 to 529.34 IU/mg. H. ulmarius laccase (376.85 IU/mg) with 1.83 fold purification had the highest specific activity of all the ligninolytic enzymes studied, followed by 2.13 fold purification in lignin peroxidase (350.57 IU/mg) and manganese peroxidase (529.34 IU/mg) with 1.77-fold purification. Three notable bands with molecular weights ranging from 43 to 68 kDa and a single prominent band with a molecular weight of 97.4 kDa were identified on a Native PAGE gel from mycelial proteins of selected mushroom fungus. The SDS PAGE profiles of the mycelial proteins from the selected mushroom fungus were similar to the native PAGE. All three partially purified ligninolytic isozymes display three bands in native gel electrophoresis, with only one prominent band in enzyme activity staining. The 43 kDa, 55 kDa, and 68 kDa protein bands correspond to laccase, lignin peroxidase, and manganese peroxidase, respectively.

Sub-acute and acute toxicity of Ferula asafoetida and Silybum marianum formulation and effect of the formulation on delaying gastric emptying.

BACKGROUND: Delayed gastric emptying play an important role in the pathology of functional dyspepsia. Owing to their functional attributes in alleviating the gastrointestinal disorders, single or polyherbal formulations have gained attention to treat the symptoms of functional dyspepsia. We have investigated the safety and efficacy of a novel formulation of Ferula asafoetida oleo resin and standardized Silybum marianum extract (Asdamarin). METHODS: The effect of asdamarin on delayed gastric emptying was investigated in Sprague Dawley rats using phenol red method. The acute and sub-acute oral toxicity was evaluated in wistar rats following OECD guidelines 425 and 407 respectively. The data were analyzed by one-way ANOVA using GraphPad Prism 5.0 software. RESULTS: Oral administration of Asdamarin dose-dependently improved the delay in gastric emptying as evident from the significant increase in the gastrointestinal transit time (p < 0.001). The LD50 of asdamarin was estimated to be more than 2000 mg/kg. Further, in the 28-day sub-acute toxicity study, the administration of 250, 500 and 1000 mg/kg of Asdamarin did not significantly altered the feed and water consuption, body weight change, biochemical and haematological parameters compared to control animals. Macroscopic and histopathological examination of vital organs revealed no toxic signs. CONCLUSION: The preliminary data from the present study provides the first evidence on the possible effectiveness of novel formulation of F. Asafoatida and S. marianum extracts in alleviating the associated symptoms of functional dyspepsia. The toxicity data indicated that Asdamarin can be considered safe up to 1000 mg/kg dose.

Potential of phloroglucinol to improve erectile dysfunction associated with streptozotocin-induced diabetes in rats.

OBJECTIVE: Diabetes is a common metabolic disease with several complications in its patients. Often, people living with diabetes develop erectile dysfunction (ED). The primary aim of this work was to investigate the effect of phloroglucinol in diabetes-induced ED in rats. METHODS: Male Wistar rats were given 52 mg/kg of streptozotocin, by intraperitoneal injection, to induce diabetes and ED. Subsequently, animals were grouped into three groups: group 1, diabetic control; group 2, low-dose phloroglucinol (150 mg/kg body weight); and group 3, high-dose phloroglucinol (250 mg/kg body weight). A group of six normal rats served as a normal control. The rats were treated with phloroglucinol for six weeks and then were assessed for treatment effects. Sexual behavior, glycosylated hemoglobin A1c (HbA1c) values, serum testosterone, serum nitric oxide (NO), blood pressure and sperm count were measured after the end of treatment. RESULTS: After a 6-week treatment period, the high dose of phloroglucinol significantly decreased HbA1c values in diabetic rats. Rats treated with phloroglucinol had increased serum testosterone, NO and sperm count. Animals treated with 250 mg/kg phloroglucinol performed similar to normal rats in the sexual behavioral study, suggesting the reversal of complications of ED. Conversely, a decrease in the blood pressure in treated groups was observed. CONCLUSION: The results highlight the protective effect of phloroglucinol in diabetes-induced ED in rats warranting further studies.

In vitro Evaluation of Antioxidant Potential of Isolated Compounds and Various Extracts of Peel of Punica granatum L.

BACKGROUND: Punica granatum L. (Lythraceae) peel has been proven to exhibit widespread pharmacological application against multitude of diseases due to the presence of bioactive principles. OBJECTIVE: The objective is to isolate the bioactive compounds from the pericarp of P. granatum and to evaluate the antioxidant activity of various extracts. MATERIALS AND METHODS: Dried peel of P. granatum was extracted with aqueous acetone and chromatographed on Diaion HP-20. Enriched fractions were rechromatographed on Sephadex LH-20 and purified on preparative high-performance liquid chromatography to identify individual compounds. The dried peel was extracted with different solvents to evaluate the antioxidant activity of the extracts. RESULTS: On the chemical investigation, three compounds were isolated and characterized as punicalagin, 2,3-(S)-hexahydroxydiphenoyl-D-glucose, and punicalin, using various spectroscopic techniques. CONCLUSION: Results indicate that the isolated compounds have possessed antioxidant activity, and aqueous, methanol, and aqueous acetone extract showed significant scavenging of 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) radicals. SUMMARY: In vitro antioxidant activity of Punica granatum extracts was evaluated by 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) assayDried peel of P. granatum was extracted with different solvents to evaluate the antioxidant activity of the extractsAqueous acetone extract was found to be most active and chromatographed further to afford punicalagin, 2,3-(S)-hexahydroxydiphenoyl-D-glucose, and punicalinThe presence of antioxidant properties of three compounds in the peel of P. granatum has been demonstrated. Abbreviations Used: HPLC: High-performance liquid chromatography; HHDP: Hexahydroxydiphenoyl; DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid); UV: Ultraviolet; PDA: Photodiode array; LC: Liquid chromatography; NMR: Nuclear magnetic resonance; MHz: Megahertz; w/v: Weight by volume; MS: Mass spectra.

Bioactive Turmerosaccharides from Curcuma longa Extract (NR-INF-02): Potential Ameliorating Effect on Osteoarthritis Pain.

BACKGROUND: Curcuma longa has long history of medicinal use in Ayurveda. A unique product NR-INF-02 was prepared from C. longa that was standardized to contain turmerosaccharides. OBJECTIVE: The present study investigated the effect of turmerosaccharides rich fraction of NR-INF-02 on monosodium iodoacetate (MIA)-induced OA pain animal model that mimics human OA. Further, the analgesic effect of turmerosaccharides rich fraction was compared to turmerosaccharides less fraction of NR-INF-02. MATERIALS AND METHODS: OA pain was chemically induced by intra-articular administration of single dose of 25 µl of 0.9% saline containing 0.3 mg MIA into the right knee of male albino Wistar rat. Turmerosaccharides rich fraction and turmerosaccharides less fraction (at 22.5, 45 and 90 mg/kg rat body weight dose levels) were administered as a single dose orally on day 5 of post-MIA injection. OA pain was measured using hind limb weight-bearing ability at 1, 3, 6, and 24 h post-test substance administration on day 5. RESULTS: Oral administration of turmerosaccharides rich fraction and turmerosaccharides less fraction (at 45 and 90 mg/kg) although significantly decreased the OA pain at all the intervals, the effect of turmerosaccharides rich fraction (57%) on OA pain was superior to turmerosaccharides less fraction (35%). CONCLUSION: Bioactive turmerosaccharides from C. longa extract contribute to the observed anti-arthritic effect in rats. SUMMARY: Osteoarthritic pain was induced by intra-articular injection of MIA into the right kneeSingle administration of TRF/TLF on day 5 resulted in dose-dependent significant reduction of OA painTRF showed better analgesic activity than TLFTRF at 45 and 90 mg/kg has similar effects on OA pain as that of tramadolTurmerosaccharides identified as bioactive constituents of C. longa extract. Abbreviations used: MIA: Monosodium iodoacetate; i.ar: Intra-articular; OA: Osteoarthritis; TRF: Turmerosaccharides rich fraction; TLF: Turmerosaccharides less fraction; PGE2: Prostaglandin E2; ROS: Reactive oxygen species.

Is Andrographis paniculata extract and andrographolide anaphylactic?

Andrographis paniculata, "King of bitters" is a popularly known medicinal plant extensively used in many parts of the world for treatment of various diseases. Since recent past, anaphylactic/allergic type adverse events were reported upon A. paniculata usage, the study aimed to evaluate the anaphylactic and anaphylactoid potential of A. paniculata extract and andrographolide (a major phytoactive of A. paniculata). The anaphylactic potential was evaluated using active systemic anaphylaxis (ASA) assay in guinea pigs. Further, the release of allergic mediators was measured in immunoglobulin E (IgE) sensitized and non-IgE sensitized Rat Basophilic Leukemia (RBL-2H3) cell lines in-vitro. A. paniculata extract or andrographolide sensitized guinea pigs following the challenge antigen administration orally and intravenously did not demonstrate any clinical signs of anaphylaxis. IgE sensitized and non- IgE sensitized RBL-2H3 cells treated with A. paniculata extract did not induce release of allergic mediators. Whereas IgE sensitized and non- IgE sensitized RBL-2H3 cells treated with andrographolide demonstrated mild to moderate release of allergic mediators. A. paniculata extract has no anaphylactic and anaphylactoid potential in in-vivo and in-vitro studies. Whereas, andrographolide effects on allergic mediators in in-vitro studies needs to be scrutinized if they are of biologically important.

Anti-stress Activity of Ocimum sanctum: Possible Effects on Hypothalamic-Pituitary-Adrenal Axis.

The present study investigated anti-stress potential of Ocimum sanctum in chronic variable stress (CVS) paradigm. Further, the possible mechanism of anti-stress was explored in vitro using cell and cell-free assays. Rats were administered O. sanctum followed by CVS regimen for a period of 16 days. On days 4, 8, 12, and 16, body weight and immobility time in forced swim test were measured. In addition, the possible inhibitory effect of O. sanctum and ursolic acid on cortisol release and CRHR1 receptor activity were studied in cell-based assays, while inhibitory effects on 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) and catechol-O-methyltransferase (COMT) were studied in cell-free assays. CVS group demonstrated less body weight gain and higher immobility time than O. sanctum administered groups, while oral administration of O. sanctum significantly increased body weight gain and decreased the immobility time. Further, O. sanctum and its constituents inhibited cortisol release and exhibited a significant CRHR1 receptor antagonist activity. Also, they had specific inhibitory activity towards 11ß-HSD1 and COMT activity. Thus, O. sanctum was found to be effective in the management of stress effects, and anti-stress activity could be due to inhibition of cortisol release, blocking CRHR1 receptor, and inhibiting 11ß-HSD1 and COMT activities. Copyright © 2016 John Wiley & Sons, Ltd.

Anti-Inflammatory Activity of Polysaccharide Fraction of Curcuma longa Extract (NR-INF-02).

The aim of the study was to investigate the safety and anti-inflammatory effects of polysaccharide fraction (F1) of Curcuma longa extract (NR-INF-02) in classical rodent models of inflammation. F1 was evaluated for its acute oral toxicity and found to be safe upto 5000 mg/kg body weight in rats. The anti-inflammatory activity of F1 was evaluated in acute (carrageenan - induced paw edema; xylene - induced ear edema) and chronic (cotton pellet - induced granuloma) models of inflammation. The results of the study demonstrated that F1 significantly (p ≤ 0.05) inhibited carrageenan-induced paw edema at 1 h and 3 h at doses of 11.25, 22.5 and 45 mg/kg body weight in rats. Also, F1 at doses of 15.75, 31.5 and 63 mg/kg significantly inhibited the xylene induced ear edema in mice. In a chronic model, F1 at 11.25, 22.5 and 45 mg/kg doses produced significant reduction of wet and dry weights of cotton pellets in rats. Overall results indicated that F1 of NR-INF-02 significantly attenuated acute and chronic inflammation in rodent models. This study emphasizes on the importance of Curcuma longa polysaccharide's role in acute and chronic inflammation.